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Partial characterization of Acanthamoeba castellanii (T4 genotype) DNase activity

Abstract

The deoxyribonuclease (DNase) activities of Acanthamoeba castellanii belonging to the T4 genotype were investigated. Using zymographic assays, the DNase activities had approximate molecular masses of 25 and 35 kDa. A. castellanii DNases exhibited activity at wide-ranging temperature of up to 60 °C and at pH ranging from 4 to 9. The DNases activities were unaffected by proteinase-K treatment, divalent cations such as Ca++, Cu++, Mg++, and Zn++, or divalent cation chelating agent ethylenediaminetetraacetic acid (EDTA) or sodium dodecyl sulfate (SDS). The non-reliance on divalent cations and homology data suggests that A. castellanii DNases belong to the class of eukaryotic lysosomal DNase II but exhibit robust properties. The DNases activity in A. castellanii interfered with the genomic DNA extraction. Extraction methods involving EDTA, SDS, and proteinase-K resulted in low yield of genomic DNA. On the other hand, these methods resulted in high yield of genomic DNA from human cells suggesting the robust nature of A. castellanii DNases that are unaffected by reagents normally used in blocking eukaryotic DNases. In contrast, the use of chaotropic agent such as guanidine thiocyanate improved the yield of genomic DNA from A. castellanii cells significantly. Further purification and characterization of Acanthamoeba DNases is needed to study their non-classic distinct properties and to determine their role in the biology, cellular differentiation, cell cycle progression, and arrest of Acanthamoeba.

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References

  1. Alsam S, Kim KS, Stins M, Rivas AO, Sissons J, Khan NA (2003) Acanthamoeba interactions with human brain microvascular endothelial cells. Microb Pathog 35:235–241

  2. Baker KP, Baron WF, Henzel WJ, Spencer SA (1998) Molecular cloning and characterization of human and murine DNase II. Gene 215:281–289

  3. Bernardi G (1971) Spleen acid deoxyribonuclease. In: Boyer PD (ed) The Enzymes. Academic Press, New York, pp 271–287

  4. Brown TJ, Cursons RTM, Keys EA (1982) Amoebae from antarctic soil and water. Appl Environ Microbiol 44(2):491–493

  5. De Jonckheere JF (1991) Ecology of Acanthamoeba. Rev Infect Dis 13(Suppl 5):S385–S387

  6. Enari M, Sakahira H, Yokoyama H, Okawa K, Iwamatsu A, Nagata S (1998) A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391:43–50

  7. Geisen S, Fiore-Donno AM, Walochnik J, Bonkowski M (2014) Acanthamoeba everywhere: high diversity of Acanthamoeba in soils. Parasitol Res 113(9):3151–3158

  8. Lepretre C, Tchakarska G, Blibech H, Lebon C, Torriglia A (2013) Apoptosis-inducing factor (AIF) and leukocyte elastase inhibitor/L-DNase II (LEI/LDNaseII), can interact to conduct caspase-independent cell death. Apoptosis 18:1048–1059

  9. Li LY, Luo X, Wang X (2001) Endonuclease G is an apoptotic DNase when released from mitochondria. Nature 412:95–99

  10. Marciano-Cabral F, Cabral G (2003) Acanthamoeba spp. as agents of disease in humans. Clin Microbiol Rev 16:273–307

  11. Martin SM, Byers TJ (1976) Acid hydrolase activity during growth and encystment in Acanthamoeba castellanii. J Protozool 23:608–613

  12. Rodriguez AM, Rodin D, Nomura H, Morton CC, Weremowicz S, Schneider MC (1997) Identification, localization, and expression of two novel human genes similar to deoxyribonuclease I. Genomics 42:507–513

  13. Siddiqui R, Khan NA (2012) Biology and pathogenesis of Acanthamoeba. Parasit Vectors 5:6

  14. Sissons J, Kim KS, Stins M, Jayasekera S, Alsam S, Khan NA (2005) Acanthamoeba castellanii induces host cell death via a phosphatidylinositol 3-kinase-dependent mechanism. Infect Immun 73:2704–2708

  15. Stins MF, Gilles F, Kim KS (1997) Selective expression of adhesion molecules on human brain microvascular endothelial cells. J Neuroimmunol 76:81–90

  16. Visvesvara GS, Moura H, Schuster FL (2007) Pathogenic and opportunistic free-living amoebae: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri, and Sappinia diploidea. FEMS Immunol Med Microbiol 50:1–26

  17. Yun SH, Seo MG, Jung BY, Kim TH, Kwon OD, Jeong KS, Rhee MH, Lee YJ, Park SJ, Kwon YS, Kwak D (2012) Characteristics of DNase activities in excretory/secretory products of infective larvae of Haemonchus contortus. J Helminthol 86:363–367

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Acknowledgments

This work was supported by the Aga Khan University.

Conflict of interest

The authors declare that they have no competing interests.

Author information

Correspondence to Naveed Ahmed Khan.

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Iqbal, J., Panjwani, S., Siddiqui, R. et al. Partial characterization of Acanthamoeba castellanii (T4 genotype) DNase activity. Parasitol Res 114, 457–463 (2015). https://doi.org/10.1007/s00436-014-4203-3

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Keywords

  • DNases
  • Acanthamoeba castellanii
  • Zymography
  • Life cycle